scholarly journals Anti-inflammatory effects of enzyme-treated asparagus extract and its constituents in hepatocytes

2016 ◽  
Vol 6 (2) ◽  
pp. 91 ◽  
Author(s):  
Mikio Nishizawa ◽  
Mana Kano ◽  
Tetsuya Okuyama ◽  
Tadayoshi Okumura ◽  
Yukinobu Ikeya
Keyword(s):  
Nitric Oxide ◽  
Heat Shock ◽  
Rat Hepatocytes ◽  
Inos Mrna ◽  
No Production ◽  
Cytokines And Chemokines ◽  
Inos Gene ◽  
Anti Inflammatory ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Background: Asparagus (Asparagus officinalis L.) is one of the most ancient vegetables, and it is rich in asparagine. Enzyme-treated asparagus extract (ETAS™; Amino Up Chemical Co., Ltd., Sapporo, Japan) is the final product of enzyme-treatment of asparagus stems and subsequent extraction. Two constituents were purified from ETAS and identified: 5-hydroxymethyl-2-furfural (HMF), an abundant constituent, and (S)-asfural, a novel constituent, which is a derivative of HMF. ETAS has been reported to increase the expression of heat shock proteins (HSPs), which are essential for the repair or removal of defective proteins. The expression of Hsp family genes is regulated by the transcription factor heat shock factor 1 (HSF1). It is unknown whether ETAS and its constituents elicit anti-inflammatory effects, such as the suppression of nitric oxide (NO), an inflammatory mediator synthesized by inducible nitric oxide synthase (iNOS) in interleukin (IL)-1β-treated hepatocytes.Objective: To examine the anti-inflammatory effects of ETAS, we treated rat hepatocytes with ETAS, or its constituents (S)-asfural or HMF, and IL-1β and then analyzed the expression of the iNOS gene and other genes involved in inflammation.Methods: Primary cultured rat hepatocytes were prepared by collagenase perfusion. ETAS, (S)-asfural, or HMF was added to the medium with IL-1β and incubated at 37 °C. When necessary, an inhibitor of HSF1 was added. NO in the medium was measured by the Griess method, and the half-maximal inhibitory concentration (IC50) values were determined. To analyze the mRNA expression, a reverse transcription-quantitative polymerase chain reaction was performed. Antibody arrays were used to determine the levels of cytokines and chemokines in the medium.Results: ETAS suppressed NO production in IL-1β-treated hepatocytes without causing cytotoxicity. ETAS decreased the levels of both iNOS mRNA and the antisense transcript, whereas it increased the levels of Hsf1 mRNA and Hsp70 mRNA. ETAS also suppressed the production of pro-inflammatory cytokines and chemokines in hepatocytes. When (S)-asfural and HMF were added to the medium, they suppressed NO production and iNOS gene expression. The IC50 value of (S)-asfural was approximately 3-fold lower than that of HMF. In contrast, (S)-asfural increased the levels of Hsf1 mRNA. Interestingly, the KRIBB11, an inhibitor of HSF1, reduced the expression of the iNOS gene. When both (S)-asfural and KRIBB11 were added, the level of iNOS mRNA was lower than when (S)-asfural alone was added.Conclusion: ETAS and its constituents (S)-asfural and HMF suppressed NO production and the expression of pro-inflammatory cytokines and chemokines, thus showing anti-inflammatory effects. Our data suggest the possibility that the increased HSF1 level is involved in suppression of NO by ETAS and its constituents, although HSF1 is essential for the expression of the iNOS gene.Keywords: nitric oxide, inducible nitric oxide synthase, inflammation, heat shock factor, asparagus

Cytokines activate inducible nitric oxide synthase gene transcription in inner medullary collecting duct cells

1995 ◽  
Vol 268 (4) ◽  
pp. F770-F777 ◽  
Author(s):  
M. G. Mohaupt ◽  
J. Schwobel ◽  
J. L. Elzie ◽  
G. S. Kannan ◽  
B. C. Kone
Keyword(s):  
Nitric Oxide ◽  
Inflammatory Cytokines ◽  
Collecting Duct ◽  
Tnf Alpha ◽  
Inos Mrna ◽  
No Production ◽  
Inos Gene ◽  
Medullary Collecting Duct ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

The effects of lipopolysaccharide (LPS) and/or inflammatory cytokines on the expression of inducible nitric oxide synthase (iNOS) were studied in mIMCD-3 cells, derived from the murine inner medullary collecting duct. Under basal conditions, the production of nitrite, a stable metabolite of NO, was negligible; however, incubation with tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IF-gamma) for 24 h resulted in a 12-fold increase in nitrite synthesis and the appearance of abundant iNOS mRNA and protein. The induction of nitrite production and iNOS mRNA was time dependent, requiring approximately 8 h for expression of significant levels of nitrite or iNOS mRNA. Coincubation with the transcription inhibitor actinomycin D or the translation inhibitor cycloheximide prevented the cytokine induction of iNOS mRNA and NO production, indicating that synthesis of intermediary proteins stimulated transcription of the iNOS gene. Nuclear run-on transcription demonstrated that the iNOS gene was transcriptionally inactive under basal conditions, but was markedly induced by TNF-alpha and IF-gamma. These results indicate that inflammatory cytokines stimulate NO production in mIMCD-3 cells by activating iNOS gene transcription in a process that requires new protein synthesis.

scholarly journals Curcumin protects liver inflammation by suppressing expression of inducible nitric oxide synthase in primary cultured rat hepatocytes

2017 ◽  
Vol 7 (9) ◽  
pp. 716 ◽  
Author(s):  
Richi Nakatake ◽  
Hidehiko Hishikawa ◽  
Hideyuki Matushima ◽  
Yusuke Nakamura ◽  
Morihiko Ishizaki ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Liver Injury ◽  
Rat Hepatocytes ◽  
No Production ◽  
Type I ◽  
Tnf Α ◽  
Cultured Rat Hepatocytes ◽  
Oxide Synthase ◽  
Necrosis Factor ◽  
Inducible Nitric Oxide

Background: Curcumin has beneficial effects on organ metabolism. However, there is little evidence that curcumin affects inflammatory mediators, such as tumor necrosis factor (TNF)-α and nitric oxide (NO). In an inflamed liver, proinflammatory cytokines stimulate liver cells, followed by the induction of inducible NO synthase (iNOS). Excessive NO produced by iNOS is one of the factors in liver injury. Therefore, inhibiting iNOS induction for preventing liver injury is important.Objective: This study aimed to investigate liver protective effects of curcumin by examining interleukin (IL)-1β-stimulated hepatocytes.Methods: Primary cultured rat hepatocytes were treated with IL-1β in the presence or absence of curcumin. Induction of NO production and iNOS, and the signaling pathway of iNOS were analyzed.Results: Simultaneous addition of IL-1β and curcumin decreased expression levels of iNOS protein and mRNA, resulting in inhibition of NO production. Curcumin also reduced mRNA expression of TNF-α and IL-6. Curcumin inhibited two essential signaling pathways for iNOS induction, NF-κB activation and type I IL-1 receptor upregulation. Transfection experiments revealed that curcumin reduced iNOS mRNA levels at the promoter activation and mRNA stabilization steps. Delayed administration of curcumin after IL-1β addition also inhibited iNOS induction.Conclusions: Curcumin affects induction of inflammatory mediators, such as iNOS and TNF-α, in part through the inhibition of NF-κB activation in hepatocytes. Curcumin may have therapeutic potential for organ injuries, including the liver.Key words: curcumin, inducible nitric oxide synthase, liver injury, primary cultured hepatocytes, nuclear factor-κB, type I interleukin-1 receptor, tumor necrosis factor-α. 

scholarly journals Anti-inflammatory Activity of Constituents Isolated from Terminalia chebula

2014 ◽  
Vol 9 (7) ◽  
pp. 1934578X1400900 ◽  
Author(s):  
Min Hye Yang ◽  
Zulfiqar Ali ◽  
Ikhlas A. Khan ◽  
Shabana I. Khan
Keyword(s):  
Nitric Oxide ◽  
Cellular Level ◽  
Inflammatory Activity ◽  
No Production ◽  
Terminalia Chebula ◽  
Arjunolic Acid ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

This study was aimed at the evaluation of the anti-inflammatory activity of twelve compounds isolated from the methanolic extract of fruits of Terminalia chebula. The activity was determined in terms of their ability to inhibit inducible nitric oxide synthase ( iNOS) and cyclooxygenase-2 (COX-2) in LPS-stimulated macrophages. Two gallotannins [chebulinic acid (1) and 2,3,6-tri- O-galloyl-β-D-glucose (2)] and two triterpenoids [arjunic acid (3) and arjunolic acid (4)] efficiently reduced nitric oxide (NO) production with IC50 values of 53.4, 55.2, 48.8, and 38.0 μM, respectively. The protein expressions of iNOS and COX-2 were decreased in macrophages by treatment with compounds 1–4 (54–69% and 33–37%, respectively) at 50 μM. This is the first report of anti-inflammatory property of 1–4 mediated by inhibition of iNOS and COX-2 activities at the cellular level.

cAMP enhances inducible nitric oxide synthase mRNA stability in cardiac myocytes

1995 ◽  
Vol 269 (6) ◽  
pp. H2044-H2050 ◽  
Author(s):  
C. V. Oddis ◽  
R. L. Simmons ◽  
B. G. Hattler ◽  
M. S. Finkel
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Cardiac Myocytes ◽  
Inducible Nitric Oxide Synthase ◽  
Mrna Stability ◽  
Neonatal Rat ◽  
Inos Mrna ◽  
No Production ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

The effects of adenosine 3',5'-cyclic monophosphate (cAMP) on cardiac myocyte nitric oxide (NO) production were studied. Maximal nitrite (NO2(-)) production by cultured neonatal rat cardiac myocytes was achieved with 500 U/ml interleukin-1 beta (IL-1 beta) for 48 h (4.6 +/- 0.3 nmol/1.25 x 10(5) cells; n = 12). Cardiac myocytes exposed to 500 U/ml IL-1 beta for 48 h stained positively for inducible nitric oxide synthase (iNOS) by immunohistochemistry. Forskolin (FSK; adenylate cyclase stimulator) or dibutyryl cAMP (DBcAMP; membrane-permeable cAMP analogue) administration alone had no effect on NO2(-) production. The addition of FSK or DBcAMP to IL-1 beta significantly increased NO2-) levels vs. IL-1 beta alone (9.7 +/- 0.6 and 10.9 +/- 0.8 vs. 4.6 +/- 0.3 nmol/1.25 x 10(5) cells per 48 h, respectively; P < 0.01; n = 12). Semiquantitative reverse transcriptase-polymerase chain reaction revealed increased iNOS mRNA in myocytes treated with FSK+IL-1 beta or DBcAMP+IL-1 beta vs. those treated with IL-1 beta alone. The addition of FSK or DBcAMP to IL-1 beta increased iNOS mRNA half-life over IL-1 beta treatment alone (10.6, 11.7 vs. 2.4 h, respectively). Cardiac myocytes do not express iNOS in response to cAMP alone. Rather, cAMP enhances iNOS mRNA stability following cytokine exposure.

Anti-Nociceptive and Anti-Inflammatory Effects of Angelicae Dahuricae Radix Through Inhibition of the Expression of Inducible Nitric Oxide Synthase and NO Production

2008 ◽  
Vol 36 (05) ◽  
pp. 913-928 ◽  
Author(s):  
Ok-Hwa Kang ◽  
Hee-Sung Chae ◽  
You-Chang Oh ◽  
Jang-Gi Choi ◽  
Young-Seob Lee ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Formalin Test ◽  
No Production ◽  
Dependent Manner ◽  
Paw Edema ◽  
Sleeping Time ◽  
Anti Inflammatory ◽  
Angelicae Dahuricae Radix ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

The extract of Angelicae Dahuricae Radix has traditionally been used as an anti-noceptive remedy in China. In this study, the methanol extract of Angelicae Dahuricae Radix (MEAD) was evaluated to determine if it has anti-noceptive and anti-inflammatory action. The anti-nociceptive activities of MEAD were evaluated by determining the writhing response and sleeping time, as well as by a formalin test. In addition, the anti-inflammatory activities of MEAD were evaluated by a vascular permeability test as well as by measuring the carrageenan-induced paw edema and conducting a myeloperoxidase (MPO) assay. MEAD (600 and 1200 mg/kg) exhibited anti-inflammatory effects on acetic acid-induced vascular permeability, carrageenan-induced paw edema, and MPO activity. Moreover, the results of the formalin test, the acetic acid-induced writhing response and the pentobarbital-induced sleeping time indicated that MEAD had anti-nociceptive effects that occurred in a concentration-dependent manner. To determine the mechanism by which MEAD exerted its effects on the expression of inducible nitric oxide synthase (iNOS) and the production of nitric oxide (NO) by treated murine macrophage RAW 264.7 cells was evaluated. Similar to the in vivo activities, both the iNOS expression and NO production were significantly suppressed by MEAD in a dose-dependent manner. Furthermore, MEAD inhibited the activating phosphorylation of ERK1/2. These results provide a scientific basis that explains the mechanism by which Angelicae Dahuricae Radix relieves inflammatory pain.

Bi-directional effects of the elevation of intracellular calcium on the expression of inducible nitric oxide synthase in J774 macrophages exposed to low and to high concentrations of endotoxin

2001 ◽  
Vol 354 (2) ◽  
pp. 351-358 ◽  
Author(s):  
Riku KORHONEN ◽  
Hannu KANKAANRANTA ◽  
Aleksi LAHTI ◽  
Mari LÄHDE ◽  
Richard G. KNOWLES ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Cell Activation ◽  
Inos Mrna ◽  
No Production ◽  
Ionophore A23187 ◽  
Inos Protein ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Nitric oxide produced through the action of inducible nitric oxide synthase (iNOS) is an important mediator in immune responses of the host. Various extracellular factors, including inflammatory stimuli, affect intracellular free Ca2+ levels ([Ca2+]i), modulating cellular signalling and gene expression. In the present study we investigated the effects of increased [Ca2+]i on NO production through the iNOS pathway in J774 macrophages. Thapsigargin (TG), a Ca2+-ATPase inhibitor, and the Ca2+ ionophore A23187 were used as tools to induce an increase in [Ca2+]i in the cytosol. This increase was confirmed by the fura 2 method. The production of NO was measured as accumulated nitrite in the cell culture medium; iNOS protein and iNOS mRNA were detected by Western blotting and reverse-transcriptase-mediated PCR respectively. The activation of nuclear factor κB (NF-κB) was investigated by electrophoretic mobility-shift assay. TG (100nM) induced a marked synthesis of iNOS mRNA, iNOS protein and NO in cells primed with a low concentration of endotoxin [lipopolysaccharide (LPS) 1ng/ml], which on its own induced barely detectable NO synthesis. Stimulation by a high concentration of LPS (100ng/ml) induced a marked expression of iNOS and NO production. Under these conditions, treatment with TG hindered the synthesis of iNOS protein and NO production by accelerating the degradation of iNOS mRNA. Treatment with TG (100nM) did not affect the NF-κB activity induced by low (1ng/ml) or high (100ng/ml) concentrations of LPS. Viability of the cells was confirmed by the 2,3-bis[2-methoxy-4-nitro-5-sulphophenyl]-2H-tetrazolium-5-carboxyaniline (‘XTT’) method; apoptosis was ruled out by propidium iodide staining and flow cytometry. A23187 (1µM) also transiently increased [Ca2+]i and had opposite effects on NO production depending on the LPS concentration. Our results show that increased [Ca2+]i induced the stimulation or suppression of NO production through iNOS in macrophages depending on the state of cell activation. These findings suggest that the receptor-mediated increase in [Ca2+]i might be an important factor in the control of the balance between the up-regulation and down-regulation of inflammatory genes, including that encoding iNOS, depending on the phase of the inflammatory response.

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